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Milk oligosaccharides are complex carbohydrates composed of various monosaccharide units linked together by glycosidic bonds. They play an essential role in promoting gut health by fostering beneficial bacteria, supporting the development of the immune system, and protecting against infections and diseases. This work compared the oligosaccharide profiles in widely utilized breeds such as Holstein and Ayrshire (Nordic Red), with the native Northern Finncattle, which is considered an endangered breed. Oligosaccharides were extracted from milk and analyzed by liquid chromatography-mass spectrometry. The composition and relative abundance of the identified oligosaccharides were characterized and compared. The statistical analyses showed that neutral, sialylated, and fucosylated oligosaccharides vary among the breeds. Ayrshire and Northern Finncattle oligosaccharides formed a cluster, while Holsteins profile shared features with both Ayrshire and Northern Finncattle. Holstein had the lowest abundance of fucosylated OS among the three breeds, with Ayrshire having the highest content followed by Northern Finncattle. The relatively higher sialylated over neutral content of Northern Finncattle is an important feature that should be preserved. Ayrshire is a good candidate to recover more diverse oligosaccharides with potential gut health implications for consumers.

There is great concern in equine sport over the potential use of pharmaceutical agents capable of editing the genome or modifying the expression of gene products. Synthetic oligonucleotides are short, single-stranded polynucleotides that represent a class of agents capable of modifying gene expression products with a high potential for abuse in horseracing. As these substances are not covered by most routine anti-doping analytical approaches, they represent an entire class of compounds that are not readily detectable. The nucleotide sequence for each oligonucleotide is highly specific, which makes targeted analysis for these agents problematic. Accordingly, we have developed a non-targeted approach to detect the presence of specific product ions that are not naturally present in ribonucleic acids. Briefly, serum samples were extracted using solid-phase extraction with a mixed-mode cartridge following the disruption of protein interactions to isolate the oligonucleotides. Following the elution and concentration steps, chromatographic separation was achieved utilizing reversed-phase liquid chromatography. Following an introduction to a Thermo Q Exactive HF mass spectrometer using electrospray ionization, analytes were detected utilizing a combination of full-scan, parallel reaction monitoring and all ion fragmentation scan modes. The limits of detection were determined along with the accuracy, precision, stability, recovery, and matrix effects using a representative 13mer oligonucleotide. Following method optimization using the 13mer oligonucleotide, the method was applied to successfully detect the presence of specific product ions in three unique oligonucleotide sequences targeting equine-specific transcripts.

Monofluoroacetate is a highly lethal toxin that causes death by inhibiting cellular adenosine triphosphate (ATP) production. The heart and brain are the primary target organs. Acute death is attributed to cardiac fibrillation and/or convulsions. Although it occurs naturally in some plants, a major source of animal intoxication is access to sodium monofluoroacetate (NaMFA) pesticide, which continues to be a concern in the USA and around the world despite restricted use in some countries including the USA. There are also concerns about misuse of this pesticide for malicious poisoning. Currently, a tissue-based diagnostic method for NaMFA intoxication in animals is lacking. There is a critical need by the veterinary diagnostic community for a simple, sensitive and reliable tissue-based diagnostic test to confirm NaMFA poisoning in animals. We have developed and extensively evaluated a sensitive novel liquid chromatography combined with tandem mass spectrometry method suitable for this purpose. The limits of detection and limits of quantitation are 1.7 and 5.0 ng/g, respectively. The accuracy and precision met or exceeded expectations. The method performance was verified using the incurred kidney obtained from animal diagnostic cases. This novel kidney-based method is now available for clinical use and can help with diagnostic purposes, including detecting potential issues related to animal foods.

Sphingolipids are an essential subset of bioactive lipids found in most eukaryotic cells that contribute to membrane biophysical properties and are involved in cellular differentiation, recognition, and mediating interactions. The described nanoHPLC-ESI-Q/ToF methodology utilizes known biosynthetic pathways, accurate mass detection, optimized collision-induced disassociation, and a robust nanoflow chromatographic separation for the analysis of intact sphingolipids found in human tissue, cells, and serum. The methodology was developed and validated with an emphasis on addressing the common issues experienced in profiling these amphipathic lipids, which are part of the glycocalyx and lipidome. The high sensitivity obtained using nanorange flow rates with robust chromatographic reproducibility over a wide range of concentrations and injection volumes results in confident identifications for profiling these low-abundant biomolecules.

Short-chain fatty acids (SCFAs) comprise the largest group of gut microbial fermentation products. While absorption of most nutrients occurs in the small intestine, indigestible dietary components, such as fiber, reach the colon and are processed by the gut microbiome to produce a wide array of metabolites that influence host physiology. Numerous studies have implicated SCFAs as key modulators of host health, such as in regulating irritable bowel syndrome (IBS). However, robust methods are still required for their detection and quantitation to meet the demands of biological studies probing the complex interplay of the gut-host-health paradigm. In this study, a sensitive, rapid-throughput, and readily expandible UHPLC-QqQ-MS platform using 2-PA derivatization was developed for the quantitation of gut-microbially derived SCFAs, related metabolites, and isotopically labeled homologues. The utility of this platform was then demonstrated by investigating the production of SCFAs in cecal contents from mice feeding studies, human fecal bioreactors, and fecal/bacterial fermentations of isotopically labeled dietary carbohydrates. Overall, the workflow proposed in this study serves as an invaluable tool for the rapidly expanding gut-microbiome and precision nutrition research field.

BACKGROUND: Effects of the renin-angiotensin-aldosterone system (RAAS) inhibitors enalapril and telmisartan on circulating RAAS in dogs with proteinuric chronic kidney disease (pCKD) are undescribed. OBJECTIVES: To characterize the RAAS in untreated dogs with pCKD compared to healthy, life-stage- and sex-matched controls, and in dogs with pCKD after 30 days of treatment with enalapril or telmisartan. ANIMALS: Dogs with pCKD (n = 36) and healthy controls (n = 20). METHODS: Retrospective study of banked samples and previously collected data. Day 0 serum equilibrium concentrations of angiotensin I, II, III, IV, 1-5, and 1-7, and aldosterone, and urinary aldosterone-to-creatinine ratio (UACR) from pCKD dogs were compared to values on day 30 of treatment with enalapril (0.5 mg/kg PO q12) or telmisartan (1 mg/kg PO q24h) and to those of healthy dogs. Data were analyzed using linear mixed models. RESULTS: Compared with healthy dogs, pCKD dogs had significantly higher Ang I, III, 1-5, and 1-7 concentrations, and UACR. Relative to pretreatment values, day 30 Ang II concentrations were significantly increased and decreased in telmisartan- and enalapril-treated pCKD dogs, respectively (both P

INTRODUCTION: While known animal exposures to human drugs of abuse (DA) were previously considered relatively uncommon in veterinary medicine, the trends are changing. Marijuana and amphetamines are among the 20 toxicants most frequently consulted about with the Pet Poison Helpline. When such exposures occur, they are typically considered emergencies. METHODS: This retrospective study describes confirmed cases of DA exposure in pets from the California Animal Health and Food Safety Laboratory System (CAHFS), 2013-2023. RESULTS: Fifty-seven samples tested positive for DA through liquid chromatography with tandem mass spectrometry analysis (qualitative method). In 75% (43/57) of the DA screen tests, the detected drugs included amphetamine-type stimulants and metabolites (methamphetamine, amphetamine, or both). In 47% (27/57) of cases, a combination of more than one drug group was found. Most cases were diagnosed from a urine specimen. In at least 32% (18/57) of cases, the samples were submitted due to suspicions of animal cruelty, and at least 41% (23/57) of the patients were deceased when the samples were submitted. DISCUSSION: More studies on the prevalence of illicit drugs in small animals, using confirmatory testing, are warranted to fully understand the significance of this emerging toxicological hazard in veterinary medicine.

Microbial competition within plant tissues affects invading pathogens' fitness. Metabolomics is a great tool for studying their biochemical interactions by identifying accumulated metabolites. Xylella fastidiosa, a Gram-negative bacterium causing Pierce's disease (PD) in grapevines, secretes various virulence factors including cell wall-degrading enzymes, adhesion proteins, and quorum-sensing molecules. These factors, along with outer membrane vesicles, contribute to its pathogenicity. Previous studies demonstrated that co-inoculating X. fastidiosa with the Paraburkholderia phytofirmans strain PsJN suppressed PD symptoms. Here, we further investigated the interaction between the phytopathogen and the endophyte by analyzing the exometabolome of wild-type X. fastidiosa and a diffusible signaling factor (DSF) mutant lacking quorum sensing, cultivated with 20% P. phytofirmans spent media. Liquid chromatography-mass spectrometry (LC-MS) and the Method for Metabolite Annotation and Gene Integration (MAGI) were used to detect and map metabolites to genomes, revealing a total of 121 metabolites, of which 25 were further investigated. These metabolites potentially relate to host adaptation, virulence, and pathogenicity. Notably, this study presents the first comprehensive profile of X. fastidiosa in the presence of a P. phytofirmans spent media. The results highlight that P. phytofirmans and the absence of functional quorum sensing affect the ratios of glutamine to glutamate (Gln:Glu) in X. fastidiosa. Additionally, two compounds with plant metabolism and growth properties, 2-aminoisobutyric acid and gibberellic acid, were downregulated when X. fastidiosa interacted with P. phytofirmans. These findings suggest that P. phytofirmans-mediated disease suppression involves modulation of the exometabolome of X. fastidiosa, impacting plant immunity.

To investigate the dissipation, terminal residues, and dietary exposure risk of flonicamid, a field trial was conducted in China, and cabbage samples were determined by modified QuEChERS pretreatment and high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The method showed good linearity (R2 ≥ 0.99), the mean recoveries ranged from 97% to 103%, with the relative standard deviation (RSD) less than 1.5%, and the matrix effect (ME) was 5%. The field trial showed that the dissipation of flonicamid in Shanxi conformed to the first-order kinetic equation with a half-life of 2.2 days. The terminal residue results showed that the residues of flonicamid in cabbage were all lower than 0.12 mg/kg. The hazard quotient (HQ) and risk quotient (RQ) were applied to dietary exposure risk assessment based on terminal residue and toxicology data. The HQ of different populations ranged from 0.17 to 0.37, and the RQ was 81.4%, indicating low risk of dietary intake of flonicamid in cabbage. [ABSTRACT FROM AUTHOR]

An applicable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method is developed to enable the efficient residue detection of fifteen acidic pesticides in water (drinking, ground, irrigating, and agricultural drainage water). The extraction and sample concentration were accomplished using the solid-phase extraction (SPE) technique. The developed method was fully validated according to the EU validation guidelines. The accuracy and precision calculations were evaluated by sample fortifications at concentrations of 0.1–1.0 µg/L. Acceptable recoveries and precisions were obtained of 70–120% and below 20%, respectively. The method showed good linearity ≥0.995. The developed method's quantitation limit was 0.1 µg/L for most studied pesticides. The uncertainty measurement, expressed as expanded uncertainty, in terms of relative standard deviation (at 95% confidence level) is within the range of ±44%. A survey study was successfully conducted by collecting 108 water samples from different water sources in Egypt during the period between June 2020 and May 2021. The obtained results showed the absence of residue for most of the acidic pesticides in the collected samples, except for bentazone which was present in 33% of the collected samples with a concentration range of 0.1 to 0.29 µg/L. [ABSTRACT FROM AUTHOR]

Persistence of spirotetramat and imidacloprid residues on tomato and onion after three applications of a combination formulation, spirotetramat 12% + imidacloprid 12%, @ 625 and 1250 mL ha−1 is reported. An easy and simple methodology was standardised for the extraction of residues in tomato, onion and soil followed by estimation and confirmation on LC-MS/MS using C18 column by keeping the positive mode of electrospray. The methodology was validated for selectivity, linearity, precision, repeatability, reproducibility and its limit of detection and quantification. The limit of quantification (LOQ) was worked out to be 0.05 mg kg−1. The results of recovery studies were consistent and more than 80% in all the matrices. No residues above LOQ were detected in tomato sample after 3rd day and after 7th day for onion samples. [ABSTRACT FROM AUTHOR]

Dithiocarbamates (DTCs) are nonsystemic, protective fungicides used all over the world to protect fruits, vegetables, and herbs from several fungal infections. Hence, it is very important to detect and quantify their residue in food. In this work, a rapid and sensitive protocol has been validated for the determination of DTCs subclasses, ethylenebisdithiocarbamate (EBDC) and propylenebisdithiocarbamate (PBDC) fungicides in fruits, vegetables, and herbs. The method was based on the decomposition of EBDC and PBDC in an alkaline medium followed by methylation into dimethyl ethylenebisdithiocarbamate (EBDC-dimethyl) and dimethyl propylenebisdithiocarbamate (PBDC-dimethyl), respectively. This was followed by applying citrate buffer acetonitrile-based QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample preparation technique. The method validation was performed according to the European quality control guidelines (SANTE/12682/2019). The precision and trueness of the method were determined from recovery experiments on six replicates of spiked blank samples at 0.01, 0.05, and 0.1 mg/kg for each commodity. The mean recoveries of EBDC-dimethyl and PBDC-dimethyl ranged from 79–119% and 74–120%, respectively. The RSD value for both analytes was in the range of 2–20. The only exception was PBDC-dimethyl in fennel matrix at 0.01 mg/kg. The limit of quantification of the method was 0.01 mg/kg with the exception of PBDC-dimethyl in fennel, which was 0.05 mg/kg. [ABSTRACT FROM AUTHOR]

Rationale: Isomerism can be an important aspect in pharmaceutical drug development. Identification of isomers can provide insights into drug pharmacology and contribute to better design of drug molecules. The general approaches to differentiate isomers include Fourier‐transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), and circular dichroism. Additionally, a commonly used method to differentiate isomers is liquid chromatography coupled with mass spectrometry (LC‐MS). Notably, LC‐MS is routinely applied to leucine and isoleucine differentiation to facilitate protein sequencing. This work focuses on isomer differentiation of widely employed thio‐succinimide structure bridging the antibody backbone and linker‐payload of antibody‐drug conjugates (ADCs). Thio‐succinimide hydrolysis stabilizes the payload‐protein structure while generating a pair of constitutional isomers: thio‐aspartyl and thio‐isoaspartyl. Methods: This paper introduces a hybrid method using ligand binding assay (LBA) and liquid chromatography coupled with tandem MS (LC–MS/MS) to reveal isomerization details of thio‐succinimide hydrolysis over time in plasma samples incubated with ADC. Application of two orthogonal dissociation methods, collision‐induced dissociation (CID) and electron‐activated dissociation (EAD) revealed different MS/MS spectra for this pair of isomers. This observation enables a unique approach in distinguishing thio‐succinimide hydrolysis isomers. Results: We observed signature [R1 + Thio + 57 + H]+, [R2 + Succ + H2O − 57 + H]+, and [R2 + Succ + H2O − 44 + 2H]2+ product ions (Succ = succinimide) that differentiated thio‐aspartyl and thio‐isoaspartyl isomers using EAD. A newly discovered [R2 + ThioSucc + H2O − 44 + 2H]2+ ion also served as additional evidence that further supported our findings. Conclusions: This study is a first‐to‐date identification of thio‐succinimide hydrolysis isomers without using synthesized reference materials. This approach should be applicable to all thio‐succinimide‐linked molecules. Correct identification of thio‐succinimide hydrolysis isomers may eventually benefit the development of ADCs in the future. [ABSTRACT FROM AUTHOR]

We report a method to determine HbA1c (glycated hemoglobin) where whole blood samples are prepared by fast hemolysis (dilution with deionized water and vortex mixing), digestion with 0.6 mg/mL endoproteinase Glu C (Glu C) in 30 mM ammonium acetate buffer (pH 4.3) at 37 °C for 45 min, and termination of the digestion by diluting with 0.1% formic acid in water, and then analysis by a gradient liquid chromatography-tandem mass spectrometry (LC–MS/MS) method with a run time of 36 s. The method is linear between 0 and 200 HbA1c/mol Hb (IFCC) with a correlation coefficient of 0.999, providing an inter-day reproducibility between 1.3 and 2.3% CV, and comparable with results from analysis of the same samples on the Roche Cobas® c 513 clinical analyzer with a correlation coefficient of 0.998. In two alternative detection workflows that were not characterized in detail, the same digested samples were purified by a magnetic bead-based solid-phase extraction (SPE) method requiring about 10 min and then analyzed using either an isocratic LC–MS/MS method or a flow injection analysis (FIA)-MS/MS method with run times of 12 s and 18 s, respectively. Our work demonstrates the feasibility of LC–MS-based methods for HbA1c determination that minimize the time required for sample preparation and measurement while preserving analytical performance and are thereby more suitable for routine clinical settings compared to traditional methods which require up to 25 h and 23 min, respectively, to prepare and measure samples. [ABSTRACT FROM AUTHOR]

Saponins encompass a diverse group of naturally occurring glycoside molecules exhibiting amphiphilic properties and a broad range of biological activities. There is a resurgence of interest in those saponins produced by marine organisms based on their potential therapeutic benefits, application in food products and most recently their potential involvement in intra- and inter-species chemical communication. The continual advancements in liquid chromatography techniques and mass spectrometry technologies have allowed for greater detection rates, as well as improved isolation and elucidation of saponins. These factors have significantly contributed to the expansion in the catalogue of known saponin structures isolated from marine invertebrates; however, there currently exists no specific chemical library resource to accelerate the discovery process. In this study, a Marine Animal Saponin Database (MASD v1.0) has been developed to serve as a valuable chemical repository for known marine saponin-related data, including chemical formula, molecular mass and biological origin of nearly 1000 secondary metabolites associated with saponins produced by marine invertebrates. We demonstrate its application with an exemplar asteroid extract (Acanthaster cf. solaris, also known as crown-of-thorns starfish; COTS), identifying saponins from the MASD v1.0 that have been previously reported from COTS, as well as 21 saponins isolated from multiple other related asteroid species. This database will help facilitate future research endeavours, aiding researchers in exploring the vast chemical diversity of saponins produced by marine organisms and providing ecological insights, and the realisation of their potential for various applications, including as pharmaceuticals. [ABSTRACT FROM AUTHOR]

Skin health relies heavily on a well-maintained cutaneous barrier. While the detrimental effects of UV radiation on the epidermis are established, the impact of blue light, a significant component of sunlight and artificial sources, is less clear. This study aims to explore blue light's influence on the reconstructed human epidermis (RHE) using two complementary analytical approaches: Raman microspectroscopy and normal phase liquid chromatography coupled with high-resolution mass spectrometry (NPLC/HR-MSn). RHE samples were exposed to blue light (415 nm and 455 nm) during different stages of their maturation. Raman spectra were acquired for both irradiated and non-irradiated (control) samples. Raman descriptors were analyzed to assess potential alterations in the structural organization of proteins and lipids' conformational changes. Additionally, lipids from RHE samples were extracted and analyzed using NPLC/HR-MSn. Blue light exposure led to changes in the structural organization of RHE lipids and proteins, as well as changes in the lipid composition. These changes varied depending on the wavelength and exposure dose. Exposure to blue light could disrupt the integrity of the skin's protective barrier, leading to increased sensitivity to environmental stressors and potential skin damage. [ABSTRACT FROM AUTHOR]

Introduction: Acute lymphoblastic leukaemia (ALL) is the most common type of childhood leukaemia with effective chemotherapeutic treatment. However, obesity has been associated with higher ALL chemoresistance rates and lower event‐free survival rates. The molecular mechanism of how obesity promotes chemotherapy resistance is not well delineated. Objectives: This study evaluated the effect of adipocyte maturation on sequestration and metabolism of chemotherapeutic drug daunorubicin (DNR). Methods: Using targeted LC‐MS/MS multi‐analyte assay, DNR sequestration and metabolism were studied in human preadipocyte and adipocyte cell lines, where expressions of DNR‐metabolizing enzymes aldo‐keto reductases (AKR) and carbonyl reductases (CBR) were also evaluated. In addition, to identify the most DNR‐metabolizing AKR/CBR isoforms, recombinant human AKR and CBR enzymes were subject to DNR metabolism. The results were further validated by AKR‐, CBR‐specific inhibitors. Results: This report shows that adipocyte maturation upregulates expressions of AKR and CBR enzymes (by 4‐ to 60‐ folds, p <.05), which is positively associated with enhanced sequestration and metabolism of DNR in adipocytes compared to preadipocytes (by ~30%, p <.05). In particular, adipocyte maturation upregulates AKR1C3 and CBR1, which are the predominate metabolic enzyme isoforms responsible for DNR biotransformation to its metabolites. Conclusion: Fat is an expandable tissue that can sequester and detoxify DNR when stimulated by obesity, likely through the upregulation of DNR‐metabolizing enzymes AKR1C3 and CBR1. Our data partially explains why obese ALL patients may be more likely to become chemoresistant towards DNR, and provides evidence for potential clinical investigation targeting obesity to reduce DNR chemoresistance. [ABSTRACT FROM AUTHOR]

Bioactive compounds in functional foods, medicinal plants and others are considered attractive value-added molecules based on their wide range of bioactivity. It is clear that an important role is occupied by polyphenol, phenolic compounds and others. Urine is an effective biofluid to evaluate and monitor alterations in homeostasis and other processes related to metabolism. The current review provides a detailed description of the formation of urine in human body, various aspects relevant to sampling and analysis of urinary metabolites before presenting recent developments leveraging on metabolite profiling of urine. For the profiling of small molecules in urine, advancement of liquid chromatography mass tandem spectrometry (LC/MS/MS), establishment of standardized chemical fragmentation libraries, computational resources, data-analysis approaches with pattern recognition tools have made it an attractive option. The profiling of urinary metabolites gives an overview of the biomarkers associated with the diet and evaluates its biological effects. Metabolic pathways such as glycolysis, tricarboxylic acid cycle, amino acid metabolism, energy metabolism, purine metabolism and others can be evaluated. Finally, a combination of metabolite profiling with chemical standardization and bioassay in functional food and medicinal plants will likely lead to the identification of new biomarkers and novel biochemical insights. [ABSTRACT FROM AUTHOR]

Chlorination of seawater forms a range of secondary oxidative species – collectively called "chlorine-produced oxidants" (CPOs) – having different biocidal, environmental and ecotoxicological properties. The chemical speciation of these compounds is an important step in attempts to assess the effectiveness of chlorination and the potential impacts of its releases. However, comprehensive determination of CPOs represents a significant analytical challenge for many reasons, including the following: CPO species are numerous, highly reactive, with short-lifetimes, difficult to isolate and generally present at low concentrations in a complex salt matrix. Literature review reveals the development of a wide variety of analytical approaches for analysis of CPOs, either collectively via group parameters or individually. A first category of these approaches was the subject of article II (also including sampling and sample preparation) of a trilogy devoted to the chemical speciation of CPOs in seawater. In this third article – which closes the trilogy – emphasis is placed on chromatographic- and mass spectrometric-based approaches. It reviews more than 80 methods, reported from 1981 to date, and thoroughly discusses their principles and performances. Methodologies involving chemical derivatization of CPOs prior to their analysis by gas or liquid chromatography coupled to mass spectrometry provide the best sensitivities, achieving sub-ppb detection limits for species for which suitable derivatization reagents are available. Online mass spectrometry approaches are attracting increasing interest for their ability to analyze multiple CPO species in real time without extensive sample preparation steps, reaching detection limits of about ppb for less polar oxidants. At the current state of metrological development, neither the methodologies based on chromatography nor those based on online mass spectrometry allow complete speciation of CPOs. Future trends and major challenges related to these approaches are discussed. [ABSTRACT FROM AUTHOR]

Background: Huoxue Tongjiang decoction (HXTJD) is an effective prescription for treating reflux esophagitis (RE). We investigated the effects of HXTJD on esophageal motility and mucosal inflammation in a rat RE model. Methods: Chemical composition of HXTJD was analyzed by ultrahigh-performance liquid chromatography Q-Orbitrap mass spectrometry (MS). The change rates of mean contraction tension forces, mean amplitudes, and mean frequencies for the lower esophageal sphincter (LES) were recorded using the isolated tissue bath system, mechanical tension transducer, and PowerLab physiological recorder. After weighing the stomach, the phenol red labeling method was used to measure the gastric emptying rate. The LES ultrastructure was observed through transmission electron microscopy. Immunofluorescence and western blotting were used to detect the number of interstitial cells of Cajal (ICC) and the expression levels of c-kit protein, connexin43 (Cx43), and stem cell factor (SCF). Flow cytometric analysis and enzyme-linked immunosorbent assay were conducted to detect the percentages of T helper 17 (Th17) cells and regulatory T (Treg) cells and the serum concentrations of interleukin 6 (IL-6), interleukin 17 (IL-17), and interleukin 10 (IL-10) in the rats. Results: We identified 28 chemical constituents in HXTJD. Regarding esophageal motility, we revealed that HXTJD increased the mean contraction tension forces, mean amplitudes, and mean frequency change rate of LES and the gastric emptying rate; decreased stomach weight; and improved the LES ultrastructure. Additionally, HXTJD increased the number of ICC-positive cells, and c-kit, Cx43, and SCF expression levels. Regarding esophageal inflammation, HXTJD significantly decreased the percentage of Th17 cells, and IL-6 and IL-17 concentrations, and increased the percentage of Treg cells and IL-10 concentration. Conclusion: HXTJD was found to be efficacious in the rat RE model. It may promote esophageal motility and alleviate the inflammatory response by activating the SCF/c-kit/ICC pathway and regulating the Th17/Treg cell balance. [ABSTRACT FROM AUTHOR]

This study examined the impact of Semen raphani on the absorption of ginsenosides from Panax ginseng C.A. Meyer (ginseng) using a Caco-2 cell model and Ultra-High-Performance Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry (UPLC-ESI-MS). Six primary ginsenosides (Rg1, Re, Rb1, Rb2, Rc, Rd) were quantified. Results showed that Semen Raphani increased the efflux rate of ginsenosides, particularly at higher concentrations, suggesting it inhibits their absorption. The research elucidates the intestinal absorption process of ginsenosides and the antagonistic mechanism of Semen Raphani against ginseng. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a common Gram-positive pathogen that causes food poisoning and foodborne illnesses. In this study, a peptide-designated NGJ1D (GAPNDLTARFIAKK) was found among 8 peptides in the fermentation broth of Lacticaseibacillus paracasei. NGJ1D had a minimum inhibitory concentration (MIC) of 62.5 μg/mL against Staphylococcus aureus and could kill the bacteria within 3 h. In a mimicked bacterial cell membrane environment, NGJ1D could change its secondary structure from a random coil to an α-helix to increase its membrane permeability and subsequent interaction with bacteria genomic DNA. Further analysis using SDS–polyacrylamide gel electrophoresis, ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS), and molecular docking revealed that NGJ1D affects the DNA-binding protein HU of S. aureus to interfere with DNA synthesis. These results indicate that novel peptides with antimicrobial potential, such as NGJ1D, could be identified in fermentation broths and can be used for application in food preservation and the prevention and treatment of infections caused by S. aureus. [ABSTRACT FROM AUTHOR]

Background: Hydroxytyrosol (HT) is a bioactive compound present in a limited number of foods such as wines, olives, and olive oils. During alcoholic fermentation, yeast converts aromatic amino acids into higher alcohols such as tyrosol, which can undergo hydroxylation into HT. The aim of this study was to validate an analytical method using ultra performance liquid chromatography coupled with mass spectrometry (UPLC/MS‐MS) to quantify HT and its precursors (tyrosine, hydroxyphenylpyruvic acid, hydroxyphenylacetaldehyde, 4‐hydroxyphenylacetic acid, and tyrosol) in wines. Their occurrence was evaluated in a total of 108 commercial Spanish wine samples. Results: The validated method simultaneously determined both HT and its precursors, with adequate limits of detection between 0.065 and 21.86 ng mL−1 and quantification limits between 0.199 and 66.27 ng mL−1 in a 5 min run. The concentration of HT in red wines was significantly higher (0.12–2.24 mg L−1) than in white wines (0.01–1.27 mg L−1). The higher the alcoholic degree, the higher was the content of HT. The bioactive 4‐hydroxyphenylacetic acid was identified in Spanish wines for the first time at 3.90–127.47 mg L−1, being present in all the samples. Conclusion: The highest HT concentrations were found in red wines and in wines with higher ethanol content. These data are useful for a further estimation of the intake of these bioactive compounds and to enlarge knowledge on chemical composition of wines. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Antidepressants remain the first-line treatment for depression. However, the factors influencing medication response are still unclear. Accumulating evidence implicates an association between alterations in gut microbiota and antidepressant response. Therefore, the aim of this study is to investigate the role of the gut microbiota-brain axis in the treatment response of venlafaxine. After chronic social defeat stress and venlafaxine treatment, mice were divided into responders and non-responders groups. We compared the composition of gut microbiota using 16 S ribosomal RNA sequencing. Meanwhile, we quantified metabolomic alterations in serum and hippocampus, as well as hippocampal neurotransmitter levels using liquid chromatography-mass spectrometry. We found that the abundances of 29 amplicon sequence variants (ASVs) were significantly altered between the responders and non-responders groups. These ASVs belonged to 8 different families, particularly Muribaculaceae. Additionally, we identified 38 and 39 differential metabolites in serum and hippocampus between the responders and non-responders groups, respectively. Lipid, amino acid, and purine metabolisms were enriched in both serum and hippocampus. In hippocampus, the concentrations of tryptophan, phenylalanine, gamma-aminobutyric acid, glutamic acid, and glutamine were increased, while the level of succinic acid was decreased in the responders group, compared with the non-responders group. Our findings suggest that the gut microbiota may play a role in the antidepressant effect of venlafaxine by modulating metabolic processes in the central and peripheral tissues. This provides a novel microbial and metabolic framework for understanding the impact of the gut microbiota-brain axis on antidepressant response. [ABSTRACT FROM AUTHOR]

To explore the efficacy and mechanisms of stimulating the nucleus accumbens (NAc) in heroin-addicted mice using focused ultrasound and microbubbles (MBs). The conditioned place preference (CPP) method was employed to establish a heroin-addicted mice model. Mice were randomized into control (C), heroin (H), heroin + ultrasound (H + U) and H + U + MBs. Ultrasound (2 MHz fundamental frequency, 1.34 MPa peak-negative pressure, 1 MHz pulse repetition frequency, 5% duty cycle, 15 min/d, over 2 d) was applied to stimulate the NAc in the latter 2 groups. Whereas H + U + MBs received an injection of sulfur hexafluoride MBs during the stimulation. Subsequently, CPP scores, open-field test (OFT), and elevated plus-maze test (EPMT) were conducted to assess behavioral changes in addiction memory, anxiety and exercise status. HE staining was performed to detect pathological structures. Neurotransmitters such as dopamine (DA), serotonin (5-HT) and glutamate (Glu) were detected using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Transmission electron microscopy (TEM) was used to observe ultrastructural changes of synapses in NAc. Immunohistochemistry (IHC) was utilized to detect Cleaved Caspase-3 in the NAc region. Western blotting (WB) was used to detect the protein expression of Cleaved Caspase-3, Bax and Bcl-2 in NAc. HE staining showed small patches of erythrocyte exudation were observed in the NAc and adjacent areas in H + U + MBs. The CPP scores of H + U + MBs were lower (p < 0.05) than H. After ultrasound treatment, all indices of the OFT and EPMT in H + U + MBs were significantly higher than H (p < 0.05). UPLC-MS/MS revealed that the levels of DA, 5-HT and Glu in H + U + MBs were lower than H (p < 0.01). TEM showed decrease the number of synapses (p < 0.05), and noticeable swelling of mitochondria, membrane damage, as well as damage to the cristae. Further detection by IHC and WB showed that the pro-apoptotic proteins Cleaved Caspase-3 and Bax increased and Bcl-2 decreased as anti-apoptotic proteins after ultrasound combined with MBs (p < 0.05). Focused ultrasound combined with MBs stimulate the NAc can weaken the addictive memory and improve anxiety of heroin-related mice. The mechanical effect of ultrasound combined with the cavitation effect may be a potential treatment for addiction. [ABSTRACT FROM AUTHOR]

With concurrent carbon and nitrogen doping, particularly in the pyrrole nitrogen configuration of Fe 2 O 3 –CuO@CN, a substantial increase in oxygen vacancy formation occurred on the catalyst surface. This phenomenon directly contributed to an increase in the rate of reactive hydroxyl radicals (•OH) generation, consequently leading to enhanced reaction efficiency during the degradation of organic pollutants. [Display omitted] • Suitable carbon and nitrogen-codoped Fe 2 O 3 –CuO composites (FeCu@CN-3) were synthesized using a convenient pyrolysis method without separation of impurities. • Oxygen vacancies, induced by carbon and nitrogen, particularly in the pyrrole nitrogen configuration bonded to iron, were instrumental in expediting the hydroxyl radicals generation rate. This phenomenon was further substantiated by electron paramagnetic resonance and density functional theory calculations. • FeCu@CN-3 exhibited strong magnetism, facilitating easy separation from the reaction solution while retaining 98% of its degradation efficiency for Methylene blue (MB) after five cycles. Additionally, the degradation rate of MB remained unaffected, even with the addition of multiple anions, maintaining a high degradation efficiency (over 80%) for various other organic pollutants. • A comprehensive mechanism underlying the degradation of MB was inferred from the high performance liquid chromatography-mass spectrometry results. Enhancing the generation of reactive hydroxyl radicals (•OH) is crucial for overcoming the limitations of the low reactivity of heterogeneous Fenton Fe-based catalysts. Researchers have explored various methods to modify catalyst structures to enhance reactivity, yet often at the expense of stability. Herein, suitable carbon and nitrogen-codoped Fe 2 O 3 –CuO composites were synthesized via pyrolysis method, demonstrating high Fenton reaction activity and remarkable stability. Experimental findings and density functional theory calculations (DFT) revealed that the presence of oxygen vacancies on the catalyst surface facilitated an increase in exposed Fe N C active sites, promoting electron transfer and the accelerating the rate of •OH generation. Moreover, carbon and nitrogen, particularly in the form of pyrrole nitrogen bonded to Fe imparted exceptional stability to the Fe N C active sites, mitigating their dissolution. Additionally, the Fe-based catalysts exhibited strong magnetism, enabling easy separation from the reaction solution while maintaining a high degradation efficiency for various organic pollutants, even in the presence of multiple anions. Furthermore, a comprehensive mechanism for methylene blue (MB) degradation was identified, enhancing the potential practical applications of these catalysts. [ABSTRACT FROM AUTHOR]

To explore independent influencing factors for clinical efficacy of roxadustat in hemodialysis patients. Hemodialysis patients treated with roxadustat were enrolled. The plasma trough concentrations of roxadustat were measured using liquid chromatography–tandem mass spectrometry (LC–MS/MS). A multiple logistic regression model was established to determine the factors that affect clinical efficacy of roxadustat in patients undergoing hemodialysis. A total of 67 hemodialysis patients were enrolled in the study. The results showed that age, blood trough concentration of roxadustat, and baseline hemoglobin (Hb) level were independent factors of clinical efficacy of roxadustat (OR = 1.06, p =.025 for age; OR = 1.001, p =.037 for plasma concentration; and OR = 0.941, p =.003 for baseline Hb), with an AUC score of 0.859. Age, blood trough concentration of roxadustat, and baseline Hb level were independent influencing factors of the response to roxadustat in hemodialysis patients. [ABSTRACT FROM AUTHOR]

To investigate the effects of canagliflozin (20 mg/kg) on Dahl salt-sensitive (DSS) rat gut microbiota and salt-sensitive hypertension-induced kidney injury and further explore its possible mechanism. Rats were fed a high-salt diet to induce hypertension and kidney injury, and physical and physiological indicators were measured afterwards. This study employed 16S rRNA sequencing technology and liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based metabolic profiling combined with advanced differential and association analyses to investigate the correlation between the microbiome and the metabolome in male DSS rats. A high-salt diet disrupted the balance of the intestinal flora and increased toxic metabolites (methyhistidines, creatinine, homocitrulline, and indoxyl sulfate), resulting in severe kidney damage. Canagliflozin contributed to reconstructing the intestinal flora of DSS rats by significantly increasing the abundance of Corynebacterium spp., Bifidobacterium spp., Facklamia spp., Lactobacillus spp., Ruminococcus spp., Blautia spp., Coprococcus spp., and Allobaculum spp. Moreover, the reconstruction of the intestinal microbiota led to significant changes in host amino acid metabolite concentrations. The concentration of uremic toxins, such as methyhistidines, creatinine, and homocitrulline, in the serum of rats was decreased by canagliflozin, which resulted in oxidative stress and renal injury alleviation. Canagliflozin may change the production of metabolites and reduce the level of uremic toxins in the blood circulation by reconstructing the intestinal flora of DSS rats fed a high-salt diet, ultimately alleviating oxidative stress and renal injury. [ABSTRACT FROM AUTHOR]

Background: This research investigates the metabolic profiles of follicular fluid (FF) samples from patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilisation and aims to identify diagnostic and therapeutic biomarkers for PCOS through lipidomic analysis. Methods: We performed non-targeted lipid analysis of FF samples from women with PCOS (n = 6) and normal controls (n = 6) using ultra-high-performance liquid chromatography-tandem mass spectrometry. Differential lipids between the two groups were screened using multidimensional statistical analysis, followed by fold change analysis and t-tests to identify potential PCOS biomarkers. Results: Multivariate statistical analysis revealed significant differences in FF lipid levels between the PCOS and control groups. Five different lipids were selected as standards, with p < .05. Phosphatidylcholine (PC), the main differentially expressed lipid, was significantly increased in the FF of the POCS group and was closely related to other lipids. Conclusions: Using ultra-high-performance liquid chromatography-tandem mass spectrometry, we investigated lipid biomarkers based on FF lipidomics to provide useful information for the discovery of diagnostic markers for PCOS. Our study identified five distinct lipids as potential markers of PCOS, with PC being the primary aberrant lipid found in the FF of patients with PCOS. [ABSTRACT FROM AUTHOR]

Optimization of the extraction process for natural compounds aims to obtain the maximum feasible yield quickly using the least amount of solvent. A single-step ultrasonic probe-assisted extraction (UPE) technique was observed to be superior compared to the exhaustive conventional and ultrasonic bath extraction for xanthophyll pigments from a hybrid genotype (DAMH-39) of African marigold (Tagetes erecta L.) in terms of less time and solvent requirement. The process variables of the UPE technique were optimized using a Box-Behnken design (BBD) of response surface methodology (RSM) followed by fine-tuning of the model using an artificial intelligence-based genetic algorithm (GA) for the maximum output of the response variable, i.e., the total xanthophyll yield. The GA-coupled RSM model predicted the optimum values of the process variables as 31.38 mL g−1 solvent-to-solid ratio, 56.56% ultrasonication amplitude, and 30 min of ultrasonication time for a maximum xanthophyll yield of 13.35 mg g−1, which was very close to the result of validation experiment (13.10 mg g−1) with a calculated error value of only 1.88%. The quality of the extracted xanthophylls by the hybrid model-optimized method was found to be superior as compared to the exhaustive techniques. However, no significant differences were observed in the antioxidant activities of the extracts obtained by different extraction techniques due to the possible interferences from the co-extractives. Analysis of the chemical constituents of the xanthophyll extract confirmed the presence of lutein linoleate, an unsaturated fatty ester of lutein for the first time in marigold along with five monoesters and three diesters of lutein, and two diesters of violaxanthin. The study signified the potential of the RSM-GA hybrid model for process optimization of extraction of natural compounds. [ABSTRACT FROM AUTHOR]

Cultivated Arachis hypogaea L., is commonly known as peanut. Its seed coat, also referred to as a shell, plays a critical role in safeguarding the seeds and facilitating the transportation of diverse metabolites. In the current work, an examination of metabolite profiling was executed utilizing UPLC-ESI–MS/MS in the seed coat (SeC), seed skin (SeS), and seed pulp (SeP) of four distinct peanut varieties of different geographical locations in India. The method was also validated according to ICH guidelines. A linear detector response was observed within a concentration range of 0.019–20.0 ng/mL, R2 = 0.999. The limit of detection (LOD) and quantification (LOQ) of the targeted compounds were found to be within the range of 0.075–0.426 and 0.227–1.292 ng/mL, respectively. The intra-day and inter-day precision demonstrated relative standard deviation %RSD < 2%. The results of Principal Component Analysis (PCA) have demonstrated that the differentiation between varieties of peanut is attributable to the existence of luteolin and rutin having a greater impact on the identification of varieties. [ABSTRACT FROM AUTHOR]

Photocatalysis is a novel approach to degrade hazardous compounds, frequently employed in environmental remediation such as eliminating methyl orange (MO) dye from wastewater. However, low efficient usage of visible light due to the large band gap of photocatalysts and its high rate of recombination limit the process. To address this issue, piezophototronics has been utilized to improve the efficacy of catalytic degradation. Specifically for this study, the piezo-photocatalytic efficiency of ZnO–MoS2 heterostructures has been realized using solar and mechanical energy in degrading MO dye. One-dimensional heterostructures with an average length of 3.34 μm and an average diameter of 872.6 nm compactly aligned on glass substrates were synthesized through a two-step hydrothermal process. Under simulated solar illumination and ultrasonic vibration, the ZnO–MoS2 effectively degraded MO, improving the degradation efficiency from 55% to 84% by introducing piezopotential in ZnO. Ultrasonication aided the photocatalysis through field-assisted separation of the photogenerated electrons and holes, reducing recombination. Coupled liquid chromatography and mass spectrometry confirmed the degradation of MO into its smaller metabolites. The catalyst films have achieved 61% degradation even after 3 times reuse. [ABSTRACT FROM AUTHOR]

Background: This study aimed to investigate the serum metabolite profiles during neoadjuvant chemoradiotherapy (NCRT) in locally advanced rectal cancer (LARC) using liquid chromatography–mass spectrometry (LC–MS) metabolomics analysis. Methods: 60 serum samples were collected from 20 patients with LARC before, during, and after radiotherapy. LC–MS metabolomics analysis was performed to identify the metabolite variations. Functional annotation was applied to discover altered metabolic pathways. The key metabolites were screened and their ability to predict sensitivity to radiotherapy was calculated using random forests and ROC curves. Results: The results showed that NCRT led to significant changes in the serum metabolite profiles. The serum metabolic profiles showed an apparent separation between different time points and different sensitivity groups. Moreover, the functional annotation showed that the differential metabolites were associated with a series of important metabolic pathways. Pre-radiotherapy (3Z,6Z)-3,6-Nonadiena and pro-radiotherapy 1-Hydroxyibuprofen showed good predictive performance in discriminating the sensitive and non-sensitive group to NCRT, with an AUC of 0.812 and 0.75, respectively. Importantly, the combination of different metabolites significantly increased the predictive ability. Conclusion: This study demonstrated the potential of LC–MS metabolomics for revealing the serum metabolite profiles during NCRT in LARC. The identified metabolites may serve as potential biomarkers and therapeutic targets for the management of this disease. Furthermore, the understanding of the affected metabolic pathways may help design more personalized therapeutic strategies for LARC patients. [ABSTRACT FROM AUTHOR]

Introduction: An inverse relationship has been identified between tacrolimus serum concentrations and donor-derived cell-free DNA (dd-cfDNA) levels after lung transplant, but limited data exists on this relationship in the kidney transplant population. Project Aim: The purpose of this evaluation was to examine the relationship between high tacrolimus variability and elevated dd-cfDNA levels in kidney and simultaneous pancreas-kidney transplant recipients at a single center. Design: Single-center, retrospective, descriptive comparative evaluation of kidney and pancreas-kidney transplant recipients who received longitudinal ddcfDNA surveillance. Intrapatient tacrolimus variability was assessed using the coefficient of variation (%CV) measured between 1 and 12 months posttransplant. Pediatrics, retransplant or multiorgan transplant recipients, and pregnant recipients were excluded. Results: One hundred fifteen recipients with 518 dd-cfDNA levels and 3028 tacrolimus troughs were assessed. Pancreas-kidney recipients had significantly higher median dd-cfDNA (0.29% vs. 0.18%, P =.034) and were excluded from analysis. Ninety-nine kidney transplant recipients were included for analysis. Recipients with tacrolimus %CV ≥30 (N = 66) had significantly higher median dd-cfDNA than %CV <30 (0.22% vs. 0.17%, P =.031). Tacrolimus %CV ≥30 demonstrated higher median peak dd-cfDNA than %CV <30, though this was not statistically significant (0.36% vs. 0.28%, P =.058). Conclusion: These data demonstrated that high intrapatient tacrolimus variability may be associated with elevated dd-cfDNA in the first year after kidney transplant. [ABSTRACT FROM AUTHOR]

An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of nobiletin and tangeretin in rat plasma, and the plasma was processed by a simple liquid-liquid extraction method with ethyl acetate. The chromatographic column was UPLC HSS T3 (50 × 2.1 mm, 1.7 μm), the mobile phase was acetonitrile-water (containing 0.1% formic acid). Multiple reaction monitoring mode (MRM) was used for quantitative analysis, nobiletin m/z 403.29 → 373.14 (cone voltage 22v, collision voltage 28v), tangeretin m/z 373.28 → 343.17 (cone voltage 20v, collision voltage 28V), tangeretin m/z 373.28 → 343.17 (cone voltage 20V, collision voltage 28V) and internal standard vitexin m/z 433.14 → 313.03 (cone voltage 32v, collision voltage 26v). The pharmacokinetics of nobiletin and tangeretin were evaluated in rats. The established UPLC-MS/MS method in the range of 2–2,000 ng mL−1 was successfully applied to the pharmacokinetics, and the calculated bioavailability of nobiletin and tangeretin was 63.9 and 46.1%, respectively. [ABSTRACT FROM AUTHOR]

A highly accurate and precise method for the simultaneous detection of 18 neonicotinoids and their metabolites in meat was developed using liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). To improve the pretreatment step of the method, five different commercially available clean-up materials (including C18+PSA (primary secondary amine), Z-Sep (with Discover DSC-C18), EMR-Lipid, SHIMSEN QuEChERS, and Clean-up LPAS) were studied in the treatment of three meat matrices: pork, duck and yellow croaker. Based on the recovery data, we found that among the five purification materials, SHIMSEN QuEChERS was slightly more effective than the others for 18 neonicotinoids. Therefore, SHIMSEN QuEChERS was used as the purification sorbent, and the extraction solvents, extraction methods and chromatographic and mass spectrometric conditions were optimized. A matrix-matched calibration method was applied for quantification. In three different meat matrixes (pork, duck, and yellow croaker), all the target compounds showed good linearity, both with values of r 2 > 0.995. The average recovery of all neonicotinoids ranges from 63.4 to 114.2% (pork), 63.0–113.2% (duck), and 63.9–110.5% (yellow croaker). Relative standard deviations were all <15% for intraday and interday precision. The values of limit of detection (LOD) and limit of quantification (LOQ) were, respectively, ranging from 0.04 to 1.0 μg kg−1 and 0.10 to 2.0 μg kg−1. Compared with previous reports, this method has advantage in LOQs, indicating that it it may be a preferred choice for the detection of neonicotinoid pesticides in meat samples. [ABSTRACT FROM AUTHOR]

The subterranean sod webworm, also known as cranberry girdler (Chrysoteuchia topiaria), is one of the most damaging insect pests in cool‐season grass grown for seed crops in Oregon. Chemical control options are limited and require irrigation or rainfall for adequate insecticide incorporation to control C. topiaria larvae. Epichloë endophytes associated with cool‐season turfgrass species and their mycotoxin profiles are well‐documented in offering plant protection against invertebrates; these fungi may offer sustainable pest management tools. Our objectives were to characterize endophyte‐mediated resistance to C. topiaria in 19 commercially available cultivars of tall fescue, perennial ryegrass, and fine fescue grown for seed in Oregon. Endophyte status (presence and viability) of fungal endophytes and their mycotoxin profiles were measured using polymerase chain reaction, and liquid chromatography‐tandem mass spectrometry, respectively. No‐choice assays were conducted in the laboratory to measure the impact of endophyte status on C. topiaria larvae in two separate no‐choice experiments. Our results suggested that increased mortality of C. topiaria larvae (R2 = 0.8526, Experiment 1; R2 = 0.6628, Experiment 2) in tall fescue cultivars was most influenced by total peramine and ergot alkaloid, and total ergoline concentrations in Experiment 1 and 2, respectively. However, no significant effect on insect mortality was found in the perennial ryegrass and fine fescue cultivars included in this study. Overall, these findings suggest a viable grass–endophyte association can be utilized as a sustainable alternative to foliar insecticides for C. topiaria management in tall fescue seed crops. Plain Language Summary: This is an applied research article. This study explored the endophyte status of 19 commercially available cultivars of three cool‐season grass species. Two experiments were conducted in which insect response was measured regarding mortality after 7–10 days of feeding on each cultivar. We found that increased mortality occurred in endophyte‐infected tall fescue cultivars. Furthermore, the insect resistance response was also found to be associated with the production of mycotoxins in these cultivars. [ABSTRACT FROM AUTHOR]

In this study, xanthine oxidase was immobilized for the first time using a novel magnetic metal–organic framework material (Fe3O4-SiO2-NH2@MnO2@ZIF-8-NH2). A ligand fishing method was established to rapidly screen XOD inhibitors from Ligusticum wallichii based on the immobilized XOD. Characterization and properties of the immobilized enzyme revealed its excellent stability and reusability. A ligand was screened from Ligusticum wallichii and identified as ligustilide by ultra-high performance liquid chromatography tandem mass spectrometry. The IC50 value of ligustilide was determined to be 27.70 ± 0.13 μM through in vitro inhibition testing. Furthermore, molecular docking verified that ligustilide could bind to amino acid residues at the active site of XOD. This study provides a rapid and effective method for the preliminary screening of XOD inhibitors from complex natural products and has great potential for further discovery of anti-hyperuricemic compounds. [ABSTRACT FROM AUTHOR]

The preparation of cellular metabolomics samples and how to achieve comprehensive coverage of different polar metabolites in cell samples in the analysis pose a challenge for cellular metabolomics. In this study, we optimized a metabolomics protocol based on ultra-high-performance liquid chromatography high-resolution mass spectrometry (UPLC/HRMS) for the extraction and detection of metabolites in A549 cells and exploration of the intervention effect of Qi-Yu-San-Long decoction (QYSLD) on A549 cells. The results indicate that the lowest level of ATP leakage was observed when A549 cells were quenched under liquid nitrogen. MeOH/chloroform/H2O (1:2:1) extraction yielded more chromatographic peaks and excellent reproducibility, and the relative extraction efficiency of most target metabolites was also high. And we optimized the chromatographic separation conditions in both HILIC and RPLC modes, enabling comprehensive detection and analysis of metabolites with varying polarities. Then, we applied the optimized method to UPLC-Q-TOF/MS-based metabolomics of A549 cells to study the mechanism of QYSLD intervention in non-small cell lung cancer (NSCLC). The CCK-8, EdU staining, and cell cycle assay showed that QYSLD inhibited the proliferation of A549 cells by interfering with the cell cycle and blocking them in the G1 phase. A total of 36 differential metabolites associated with the antitumor effects of QYSLD on NSCLC were identified, mainly involving nicotinate and nicotinamide metabolism, sphingolipid metabolism, and glycerophospholipid metabolism. And western blotting confirmed that the change in 1-methylnicotinamide levels after QYSLD intervention was associated with the inhibition of nicotinamide N-methyltransferase expression in A549 cells. [ABSTRACT FROM AUTHOR]

Higher serum cortisol levels appear to be associated with stress that can overlap or manifest anxiety, fatigue, depression, and sleep dysfunction. These are common and intrusive non-motor symptoms of Parkinson's disease (PD). Thus, stress has been proposed to mediate Parkinson's disease development, and cortisol has been suggested as a biomarker for the generation of stress-related symptoms in Parkinson's disease. This study describes sensitive and robust disposable pipette extraction (DPX) and ultra-efficient liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method to determine cortisol and cortisone (as potential endocrine biomarkers for Parkinson's disease) in 24-h urine and saliva samples obtained from Parkinson's disease patients. Important parameters on DPX extraction were optimized to achieve the best extraction recovery and cleanup efficiency. The proposed method was linear from 0.5 (lower limit of quantification) to 500 ng mL−1 for cortisol and from 3.0 (lower limit of quantification) to 500 ng mL−1 for cortisone. To determine whether urinary cortisol and urinary cortisone are adequate as biomarkers to evaluate the level of anxiety in patients suffering from Parkinson's disease, twenty-nine Parkinson's disease patients (18 with anxiety and 11 without anxiety) were selected for urine analysis. Based on the obtained results, 24-h urine samples obtained from Parkinson's disease patients with anxiety had higher cortisone levels than samples obtained from healthy controls. Receiving operating curves (ROC) analysis, which presented the area under the ROC curve (AUC = 0.733), showed that urinary cortisone levels (µg/24-h urine) were sensitive (56.3%) and specific (93.3%) for distinguishing Parkinson's disease patients with anxiety from healthy controls. In terms of salivary results, PD patients' samples taken 30 min after waking up had higher cortisol and cortisone levels than healthy controls, while their samples taken at night had lower cortisol and cortisone levels. [ABSTRACT FROM AUTHOR]

Per- and polyfluoroalkyl substances (PFAS) are widely used in industry, residential, and consumer products. Studies have shown associations between high PFAS exposure and adverse health effects. In 2022, the National Academies of Science, Engineering, and Medicine (NASEM) published Guidance on PFAS Exposure, Testing, and Clinical Follow-up providing laboratory and clinical direction. The Guidance suggests nine PFAS should be measured in serum or plasma specimens and summed to provide a total PFAS concentration using a NASEM-recommended method. Follow-up clinical recommendations are based on the calculated PFAS NASEM summation. We developed and validated a liquid chromatography-tandem mass spectrometry (LC–MS/MS) method in accordance with NASEM recommendations but distinguished by the ability to separate closely related structural isomers. As part of our validation, PFAS prevalence was evaluated in a population survey comprised of clinical donor and remnant specimens (n = 1023 in total). In this study, 82.2% of the specimens had PFAS NASEM summations of 2 to < 20 ng/mL and 2.5% had a summation ≥ 20 ng/mL. The median PFAS NASEM summation was 4.65 ng/mL in this study, lower than the 7.74 ng/mL median observed in the 2017–2020 Centers for Disease Control and Prevention, National Health and Nutrition Examination Survey (n = 3072). This lower median PFAS NASEM summation may reflect a decline in PFAS population levels over time or sample population exposure differences. [ABSTRACT FROM AUTHOR]

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants with bioaccumulation potential, particularly affecting aquatic ecosystems and human health also via fish consumption. There is therefore a need for reliable extraction methods and studies to accurately assess PFAS levels in fish, crucial for understanding bioaccumulation and potential toxicological effects on both fish and humans through consumption. This study investigated PFAS levels in freshwater fish from Swiss lakes, focusing on six common species: Coregonus wartmanni, Cyprinus carpio, Oncorhynchus mykiss, Perca fluviatilis, Salmo trutta, and Squalius cephalus. Utilizing an optimized QuEChERS extraction method, 15 PFAS were analyzed in 218 fish fillet samples using liquid chromatography-mass spectrometry (LC–MS/MS). The results were compared to EU regulations and EFSA guidelines for tolerable weekly intake (TWI), with a specific focus on correlations between fish size and PFAS concentration. Our findings reveal significant PFAS contamination, particularly in Perca fluviatilis with perfluorooctane sulfonic acid (PFOS) and perfluorohexane sulfonic acid (PFHxS) levels often exceeding EU safety limits. TWI, calculated for a person of 70 kg body weight and an intake of 200 g of fish fillet, is exceeded in 95% of Coregonus wartmanni, 100% of Squalius cephalus, and in 55%, 50%, and 36% of the specimens Oncorhynchus mykiss, Salmo trutta, and Perca fluviatilis respectively. Correlation analysis between PFAS concentration and fish size in 121 Salmo trutta specimens revealed significant positive correlations for perfluorobutane sulfonic acid (PFBS), perfluorodecanoic acid (PFDA), and perfluorohexane sulfonic acid (PFHxS), and a negative correlation for perfluoropentanoic acid (PFPeA). These results underscore the critical need for continuous monitoring and regulatory efforts to mitigate PFAS exposure risks to both ecosystems and human health. [ABSTRACT FROM AUTHOR]

We explored the association between aerobic capacity (AC) and the skeletal muscle proteome of McArdle (n = 10) and wild‐type (n = 8) mice, as models of intrinsically ‘low’ and ‘normal’ AC, respectively. AC was determined as total distance achieved in treadmill running until exhaustion. The quadriceps muscle proteome was studied using liquid chromatography with tandem mass spectrometry, with the Search Tool for the Retrieval of Interacting Genes/Proteins database used to generate protein–protein interaction (PPI) networks and enrichment analyses. AC was significantly associated (P‐values ranging from 0.0002 to 0.049) with 73 (McArdle) and 61 (wild‐type) proteins (r‐values from −0.90 to 0.94). These proteins were connected in PPI networks that enriched biological processes involved in skeletal muscle structure/function in both groups (false discovery rate <0.05). In McArdle mice, the proteins associated with AC were involved in skeletal muscle fibre differentiation/development, lipid oxidation, mitochondrial function and calcium homeostasis, whereas in wild‐type animals AC‐associated proteins were related to cytoskeleton structure (intermediate filaments), cell cycle regulation and endocytic trafficking. Two proteins (WEE2, THYG) were associated with AC (negatively and positively, respectively) in both groups. Only 14 of the 132 proteins (∼11%) associated with AC in McArdle or wild‐type mice were also associated with those previously reported to be modified by aerobic training in these mice, providing preliminary evidence for a large divergence in the muscle proteome signature linked to aerobic training or AC, irrespective of AC (intrinsically low or normal) levels. Our findings might help to gain insight into the molecular mechanisms underlying AC at the muscle tissue level. [ABSTRACT FROM AUTHOR]

Background: Headache is a pain disorder present in populations world-wide with a higher incidence in females. Specifically, the incidences of medication overuse headache (MOH) have increased worldwide. Comorbidities of MOH include photosensitivity, anxiety, "brain fog", and decreased physical activity. The FDA-approved long-lasting selective β2-adrenergic receptor agonist, formoterol, is currently approved for use in severe asthma and chronic obstructive pulmonary disease. Recently, interest in repurposing formoterol for use in other disorders including Alzheimer's disease, and neuropathic pain after spinal cord injury and traumatic brain injury has gained traction. Thus, revisiting known side-effects of formoterol, like headache and anxiety, could inform treatment paradigms. The endocannabinoid (eCB) system is implicated in the etiology of preclinical headache, with observed decreases in the circulating levels of endogenous cannabinoids, referred to as Clinical Endocannabinoid Deficiency. As cross-talk between the eCB system and adrenergic receptors has been reported, this study investigated the role of the eCB system and ability of formoterol to induce headache-like periorbital allodynic behavior. Methods: Female 8-week-old C57Bl/6J mice were treated daily with formoterol (0.3 mg/kg, i.p.) for up to 42-days, during which they were assessed for periorbital allodynia, open field/novel object recognition, and photosensitivity. At the end of the study, the periaqueductal grey (PAG), a brain region known to contribute to both headache induction and maintenance, was collected and subjected to LC-MS to quantify endocannabinoid levels. Results: Mice exhibited periorbital allodynia at nearly all time points tested and photosensitivity from 28-days onward. Levels of endocannabinoids, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), along with cannabinoid receptor 1 (CB1R) expression were altered by both age and upon treatment with formoterol. Administration of FAAH/MAGL inhibitors, to target the eCB system, and a non-selective cannabinoid receptor agonist, WIN 55,212 reversed the formoterol-induced periorbital allodynia. Conclusions: These results suggest that formoterol is dysregulates eCB tone to drive headache-like periorbital allodynic behaviors. These results could help inform preventative treatment options for individuals receiving formoterol, as well as provide information on the interaction between the eCB and adrenergic system. [ABSTRACT FROM AUTHOR]

Mycoplasma synoviae is an important avian pathogen that causes respiratory infections and arthritis symptoms in chickens and turkeys, resulting in significant economic damage to the poultry farming industry worldwide. Cell adhesion is a vital stage of Mycoplasma infection, and the proteins associated with this process play an important role in its pathogenesis. Elongation factor thermo stable (EF-Ts) is an important factor in prokaryotic biosynthesis that serves as a guanosine exchange factor for elongation factor thermo unstable (EF-Tu). To date, little is known about the role of EF-Ts in Mycoplasma infection. In this study, we identified EF-Ts as an immunogenic protein in M. synoviae through liquid chromatography with tandem mass spectrometry (LC–MS/MS) screening. We constructed an E. coli recombinant expression vector and prepared a highly efficient rabbit antiserum. Immunoblot analysis and suspension immunofluorescence revealed that the EF-Ts is located in both the cell membrane and cytoplasm. The prepared rabbit EF-Ts antiserum exhibited complement-dependent Mycoplasma-killing activity and inhibited the adhesion of rEF-Ts and M. synoviae to DF-1 cells. An in-vitro binding assay showed that EF-Ts could bind to fibronectin (Fn) and chicken plasminogen (cPlg) in a dose-dependent manner. In addition, EF-Ts could internalize into cells through lipid rafts and clathrin-dependent endocytosis and induce DF-1 cell proliferation. In conclusion, our studies demonstrated that MS EF-Ts is a potentially immunogenic, novel adhesion protein that acts as a critical virulence factor in M. synoviae adhesion to host cells during infection. These studies further deepen our understanding of the pathogenic mechanism of M. synoviae. [ABSTRACT FROM AUTHOR]

Background: Research has demonstrated that obesity may be associated with rheumatoid arthritis (RA). In addition, Dysbiosis of intestinal microbiota and their metabolites has been linked to the occurrence and development of RA and obesity. However, the mechanism by which obesity affects RA remains unclear.In this study, we explored the impact of high fat diet(HFD) on collagen-induced arthritis (CIA) rats and revealed its mechanisms based on gut microbiota and metabolomics. Methods: Based on diet and modeling, rats were divided into normal group (Con), CIA model group, HFD group (HFD), and HFD + CIA group (HCIA). The effect of HFD on arthritis in CIA rats were investigated based on the arthritis index (AI), weight, blood lipid levels, and inflammatory cytokines. Moreover, HE staining and micro-CT were performed to evaluated the effect of HFD on the pathology of joints and synovial tissues in CIA rats.16S rRNA amplicon sequencing and liquid chromatography-mass spectrometry (LC–MS) were employed to explore changes in gut microbiota and short-chain fatty acids (SCFAs). Results: The AI scores, inflammatory cytokines and bone destruction parameters in the HCIA group were significantly higher than those in the other three groups. The results of 16S rRNA amplicon sequencing and metabolomics showed that compared with the other three groups, the expression of g_Muribaculaceae and butyric acid were reduced in the HCIA group. Spearman and linear correlation analyses revealed a positive correlation between g_Muribaculaceae abundance and butyric acid levels. Conclusions: HFD stimulated butyric acid metabolism dysbiosis, altered microbiota, and aggravated inflammatory response in CIA rats. [ABSTRACT FROM AUTHOR]

Background: Zearalenone (ZEN), a secondary metabolite of Fusarium fungi, is one of the most common mycotoxins in global food supplies such as cereal grains and processed food. ZEN and its metabolites are commonly referred to as mycoestrogens, due to their ability to directly bind nuclear estrogen receptors α (ER-α) and β (ER-β). Zeranol, a synthetic mycoestrogen, is administered to U.S. cattle as a growth promoter. Despite widespread human exposure and ample evidence of adverse reproductive impacts in vitro and in vivo, there has been little epidemiological research on the health impacts of ZEN exposure during pregnancy. The objective of our study was to examine associations between ZEN and gestational weight gain (GWG). Methods: Urine samples were collected in each trimester from pregnant participants in the UPSIDE cohort (n = 286, Rochester, NY, USA). High performance liquid chromatography and high-resolution tandem mass spectrometry were used to quantify concentrations of ZEN as well as ∑mycoestrogens (composite sum of ZEN metabolites; ng/ml). Maternal weights at clinical visits were abstracted from medical records. We fitted longitudinal models of specific-gravity adjusted, log-transformed ZEN and ∑mycoestrogens in relation to total GWG (kilograms) and GWG rate (kilograms/week). We additionally examined risk of excessive GWG (in relation to Institute of Medicine guidelines) and considered effect modification by fetal sex. Results: ZEN and ∑mycoestrogens were detected in > 93% and > 95% of samples, respectively. Mycoestrogen concentrations were positively associated with total GWG (ZEN β:0.50 kg; 95%CI: 0.13, 0.87) and GWG rate (ZEN β:0.20 kg/week; 95%CI: 0.01, 0.03). Associations tended to be stronger among participants carrying male (versus female) fetuses and results were robust to adjustment for diet. Conclusions: Mycoestrogen exposure during pregnancy may contribute to greater GWG. Future research is needed to understand potential influences on downstream maternal and offspring health. [ABSTRACT FROM AUTHOR]

Background: The brown planthopper (BPH) is an economically significant pest of rice. Bph3 is a key BPH resistance gene. However, the proteomic response of rice to BPH infestation, both in the presence and absence of Bph3, remains largely unexplored. Results: In this study, we employed tandem mass tag labeling in conjunction with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis to identify differentially expressed proteins (DEPs) in rice samples. We detected 265 and 125 DEPs via comparison of samples infected with BPH for 2 and 4 days with untreated samples of the BPH-sensitive line R582. For the Bph3 introgression line R373, we identified 29 and 94 DEPs in the same comparisons. Bioinformatic analysis revealed that Bph3 significantly influences the abundance of proteins associated with metabolic pathways, secondary metabolite biosynthesis, microbial metabolism in diverse environments, and phenylpropanoid biosynthesis. Moreover, Bph3 regulates the activity of proteins involved in the calcium signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, and plant hormone signal transduction. Conclusions: Our results indicate that Bph3 enhances the resistance of rice to BPH mainly by inhibiting the down-regulation of proteins associated with metabolic pathways; calcium signaling, the MAPK signaling pathway, and plant hormone signal transduction might also be involved in BPH resistance induced by Bph3. [ABSTRACT FROM AUTHOR]

Background: Chrysanthemi Flos is a traditional Chinese medicine with a long history of medicinal use. Prior research suggests that the intrinsic composition of Chrysanthemi Flos is affected by shade-drying and oven-drying methods. Nevertheless, the effects of these methods on the proteins and metabolites of Chrysanthemi Flos have not been extensively studied. Methods: The TMT (tandem mass tag) quantitative proteomics method and the LC–MS/MS (liquid chromatography-tandem mass spectrometry) non-targeted metabolomics method were used to systematically study the differences in the proteins and metabolites during the process of drying Chrysanthemi Flos in the shade and an oven. Results: Differentially accumulated metabolites and abundant proteins were primarily enriched in the purine metabolism, pyrimidine metabolism, cyanogenic amino acid metabolism, phenylpropanoid biosynthesis, and starch and sucrose metabolism pathways. Primary metabolites, such as guanine, xanthine, cytidine 5'-diphosphate serine, L-isoleucine, stearidonic acid, alginate, and inulin, play a crucial role in providing energy for Chrysanthemi Flos to withstand desiccation stress. The upregulation of ferulate-5- hydroxylase (F5H), shikimate O hydroxycinnamoyltransferase (HCT), caffeoyl-CoA O-methyltransferase (CCoAOMT), and chalcone isomerase (CHI) enzymes promotes the synthesis of flavonoids, including sinapic acid, caffeoyl shikimic acid, and naringenin chalcone, which possess antioxidant properties. Despite the notable improvements in energy metabolism and antioxidant capacity, these enhancements proved insufficient in halting the senescence and ultimate demise of Chrysanthemi Flos. Moreover, the shade-drying method can inhibit protein expression and promote the accumulation of bioactive components, but the drying efficiency is low, while the oven-drying method exhibits rapid drying efficiency, it does not effectively preserve the components. Conclusion: Our study offers a comprehensive explanation for the changes in protein expression and metabolite conversion observed in shade-dried and oven-dried Chrysanthemi Flos, also providing a foundation for optimizing the drying process of Chrysanthemi Flos. [ABSTRACT FROM AUTHOR]

Objective: To assess the mechanism of supraspinatus tendinopathy induced by type 2 diabetes mellitus (T2DM) in rats using untargeted metabolomics analysis. Methods: The liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics approach was used to screen tendon biomarkers of supraspinatus tendinopathy in rats with T2DM. Seventy-eight Sprague-Dawley rats were divided into normal group (NG) and T2DM groups. Rats in T2DM groups were divided into 12-week (T2DM-12w), and 24-week (T2DM-24w) subgroups according to the time point of the establishment of the T2DM rat model. Histological evaluation (modified Bonar score) and biomechanical testing were used to analyze the adverse effects of type 2 diabetes on the tendon of the supraspinatus muscle in rats.Three comparable groups were set up, including T2DM-12w group vs. NG, T2DM-24w group vs. NG, and T2DM-24w group vs. T2DM-12w group. Differentially expressed metabolites (DEMs) in the supraspinatus tendons in the three groups of rats were analyzed using LC-MS, and data were analyzed using multivariate statistical methods to screen potential biomarkers. The DEMs included in the intersection of the three groups were identified as those associated with the development of diabetic supraspinatus tendinopathy, and trend analysis and pathway topology analysis were performed. Results: With the progression of diabetes, the tendinopathy of the supracinatus muscle of diabetic rats gradually intensified, mainly manifested as inflammatory reactions, disordered collagen fibers, fat infiltration, and increased modified Bonar score. The intersection of DEMs among the three comparable groups was resulted in the identification of 10 key DEMs, in which melezitose and raffinose showed a continuous increasing trend with the prolongation of disease course. By pathway topology analysis, 10 DEMs (P < 0.01) were mainly associated with the pathways of galactose metabolism, which could be involved in the development of diabetes-induced supraspinatus tendinopathy. Conclusion: T2DM causes tendinopathy of the supraspinatus muscle in rats. 10 key DEMs obtained by untargeted metabolomics assay suggested that the development of diabetes-induced supraspinatus tendinopathy was associated with changes in metabolic pathways, such as galactose metabolism. melezitose and raffinose hold promise as a biomarker for disease discrimination and/or disease indication in diabetic supraspinatus tendinopathy. [ABSTRACT FROM AUTHOR]